JP2003286358A - Substrate for microchip and method for manufacturing the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a substrate for a microchip which can hold in a high density a functional group fixating a biochemically active substance and which can be manufactured by a simplified cheap method, as well as to provide a manufacturing method thereof. <P>SOLUTION: The substrate for a microchip, which is used for fixating the biochemically active substance, is characterized in that on the surface of a saturated cyclic polyolefin resin using as the raw material various monomers including norbornene or a norbornene derivative there is formed a layer 500 Åthick or thinner consisting of a silicon compound. This substrate can be manufactured by the dry coating of silicon oxide by vacuum deposition method, sputtering method, CVD method, and the like. <P>COPYRIGHT: (C)2004,JPO

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a microchip substrate which serves as a support for immobilizing a biochemically active substance and a method for producing the same.

[0002]

2. Description of the Related Art At present, glass substrates are the mainstream for microchip substrates such as protein chips and DNA chips. The reason is that the characteristics of the microchip are that the background is low when the detection target is measured with a fluorescent label, and that it is excellent in heat resistance and solvent resistance. However, glass has drawbacks that it is easily broken, surface treatment is relatively difficult, and mass production of substrates having excellent surface precision is difficult. Therefore, plastic is drawing attention as a material that replaces glass. Plastic is considered to be widely used in the future as a material for substrates for microchips because it is inexpensive and allows mass production of homogeneous products by molding. Functional groups are required to immobilize biochemically active substances such as proteins on the surface of plastic substrates. As a method for obtaining a substrate into which a functional group such as an amino group or an aldehyde group is introduced, a method of molding the substrate with a copolymer containing a monomer having a functional group, or under a gas atmosphere containing oxygen or oxygen There is a method of performing a surface treatment using a carboxyl group or a hydroxyl group introduced by an oxidation treatment such as a low temperature plasma discharge treatment. When synthesizing a copolymer having a functional group for immobilizing a physiologically active substance, a small amount of plastic is required to make it for the purpose of using only a microchip substrate, and is a much smaller lot than a general-purpose plastic. Since the production is done at, the manufacturing cost becomes high. On the other hand, when an oxidation treatment such as a low temperature plasma discharge treatment is performed,
Carboxyl groups and hydroxyl groups are not introduced into all the carbon molecules on the surface, and it is still necessary to improve the introduced amount of functional groups.

[0003]

DISCLOSURE OF THE INVENTION In order to solve the above-mentioned problems when a plastic substrate is used instead of the glass substrate, the present invention holds functional groups for immobilizing biochemically active substances at a high density. It is an object of the present invention to provide a plastic microchip substrate that can be manufactured by a simple and inexpensive method and can replace glass, and a manufacturing method thereof.

[0004]

As a result of studies to solve the above-mentioned problems, the present inventors have found that a vacuum of silicon oxide is applied to a norbornene resin support as one of the processing steps for introducing various functional groups. By vapor deposition, a high-density hydroxyl group was introduced on the surface to complete a microchip substrate. That is, the present invention provides: (1) a microchip substrate characterized in that a layer made of a silicon compound is formed on at least a part of a surface of a saturated cyclic polyolefin resin, (2) a saturated cyclic polyolefin resin and norbornene Alternatively, the microchip substrate according to (1), which contains at least one norbornene derivative as a monomer unit, (3) the silicon compound layer forming the surface of the support has a thickness of 500 Å or less, ) Or (2), the microchip substrate, (4) a method for producing a microchip substrate, characterized in that a layer made of a silicon compound is formed on the surface of a saturated cyclic polyolefin resin by a dry coating method. (5) Dry coating method is vacuum deposition method, sputtering method, CV
The method for producing a microchip substrate according to (4), which is any one of the D methods.

[0005]

BEST MODE FOR CARRYING OUT THE INVENTION The saturated cyclic polyolefin-based resin used in the present invention has a low background when measuring an object to be detected with a fluorescent label as a substrate for microchips, and is excellent in heat resistance and solvent resistance. There is. The specific molecular structure is a polymer having a cyclic olefin structure alone or a saturated polymer obtained by hydrogenating a copolymer of a cyclic olefin and an α-olefin, and is preferably represented by the general formula (1). And a hydrogenated product of a ring-opening polymer of norbornene.

[0006]

[Chemical 1] (However, in the above formula (1), R ₁ and R ₂ may be the same or different from each other as hydrogen or a hydrocarbon residue having 1 to 10 carbon atoms, and R ₁ and R ₂ form a ring with each other. The polymer having a structural unit represented by the general formula (1) is norbornene as a monomer and an alkyl or alkylidene-substituted product thereof, specifically, 5-methyl-2-norbornene, 5, 6-dimethyl-2-norbornene, 5
-Ethylidene-2-norbornene and the like, and ring opening obtained by ring-opening polymerization using dicyclopentadiene, 2,3-dihydrodicyclopentadiene, and their alkyl-substituted compounds such as methyl and ethyl. There is a saturated polymer produced by hydrogenating the polymer.

Further, a polymer of a cyclic olefinic monomer represented by the general formula (2), or ethylene, propylene, isopropylene, 1-butene, 3-methyl-1-butene, 1-pentene, 1-hexene, etc. A saturated polymer produced by hydrogenating a random copolymer of α-olefin and the cyclic olefin-based monomer represented by the general formula (2) may be used.

[0008]

[Chemical 2] (However, in the above formula (2), R ₁ and R ₈ are selected from the group consisting of hydrogen, a halogen atom and a hydrocarbon residue;
_{5 to} R ₈ may form a ring with each other. )

Further, a polymer of a cyclic olefin monomer represented by the general formula (3), or ethylene, propylene, isopropylene, 1-butene, 3-methyl-1-
A saturated polymer produced by hydrogenating a random copolymer of an α-olefin such as butene, 1-pentene, 1-hexene and the cyclic olefin monomer represented by the general formula (3) may be used.

[0010]

[Chemical 3] (However, in the above formula (2), R ₁ and R ₁₂ are selected from the group consisting of hydrogen, a halogen atom and a hydrocarbon residue,
R _{9 to} R ₁₂ may form a ring with each other. )

A base material which is a support for immobilization of a biochemically active substance is formed by using a saturated polyolefin resin represented by the general formulas (1) to (3) as a raw material. Is not particularly limited. In consideration of moldability, methods such as extrusion molding, compression molding, and injection molding are suitable, and in consideration of the test method and handling, a film sheet,
It is preferable to form beads, slide plates and the like into the shape of the base material.

Next, these substrates are dry-coated with silicon oxide to introduce a layer of a silicon compound having a thickness of 500 Å or less on the surface of the substrates. If the thickness of the silicon compound layer exceeds 500Å, cracks may occur on the surface when a bending stress is applied to the microchip substrate, and the surface uniformity and flatness and other shape characteristics are not impaired. It cannot be used as a substrate.

The method of dry coating is not particularly limited, but the vacuum deposition method, the sputtering method, and the CVD method are simple and not limited to the shape, and the amount of the silicon compound introduced is easily controlled, which is preferable. It is a vacuum deposition method. By the method described above, a hydroxyl group is introduced for each silicon atom present on the surface of this support in an aqueous solution, and more hydroxyl groups are introduced than in the case where treatment is performed by another method. It is possible to introduce a high density of functional groups required for immobilization of.

[0014]

EXAMPLES Next, examples of the present invention will be described. Example 1 A microchip substrate (76 mm × 26 mm × 1 mm) was injection-molded using a hydrogenated product of a random copolymer of ethylene and dicyclopentadiene. Then, the vapor deposition source made of silicon monoxide is irradiated with an electron gun under reduced pressure and heated to generate vapor of silicon oxide.
It was vacuum-deposited so as to be 50Å.

Comparative Example 1 A microchip substrate (76 mm × 26 mm × 1 mm) was injection-molded using a hydrogenated product of a random copolymer of ethylene and dicyclopentadiene. After that, high-frequency low-temperature plasma was generated while oxygen gas was passed under a reduced pressure to perform an oxidation treatment for 10 minutes.

Comparative Example 2 A microchip substrate (76 mm × 26 mm × 1 mm) made of quartz glass was used.

The microchip substrates of Examples and Comparative Examples were immersed in a 5% methanol solution of a primary aminosilane coupling agent and left for 2 hours for aminosilane treatment.
This was immersed in a 5% aqueous solution of glutaraldehyde to introduce an aldehyde group. 0.1 μL of a solution prepared by adjusting biotin hydrazide to 0.1 mg / ml is spotted on these and left to stand at 37 ° C. and 99% humidity for 120 minutes to immobilize biotin hydrazide on the substrate. Next, unfixed biotin hydrazide is removed by washing, and the substrate is immersed in a solution prepared by adding 1% by weight of skim milk and left at 37 ° C. for 90 minutes for blocking treatment. Unfixed skim milk is washed away, immersed in a solution of rhodamine-labeled avidin adjusted to 1 μg / ml, and left at 37 ° C. for 30 minutes to bind biotin and rhodamine-labeled avidin. The unfixed rhodamine-labeled avidin was removed by washing, and the fluorescence intensity was measured at an excitation wavelength of 488 nm and a measurement wavelength of 515 nm for comparative evaluation. The evaluation results are shown in Table 1. From this, it was shown that the above-mentioned Example has a higher biotin adsorption capacity than the method of Comparative Example 1 and is equivalent to the glass substrate of Comparative Example 2.

[0018]

[Table 1]

[0019]

Industrial Applicability The microchip substrate of the present invention, in which a silicon compound layer is present on the surface of a saturated cyclic polyolefin resin, is capable of holding functional groups for immobilizing biochemically active substances at a high density, simple and inexpensive. Can be manufactured by various methods.

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Claims (5)

[Claims] 1. A substrate for a microchip, which is a substrate molded from a saturated cyclic polyolefin resin and has a layer made of a silicon compound formed on at least a part of its surface. 2. The substrate for a microchip according to claim 1, wherein the saturated cyclic polyolefin resin contains at least one of norbornene or a norbornene derivative as a monomer unit. 3. A layer made of a silicon compound has a thickness of 500Å
The substrate for microchip according to claim 1 or 2, which is as follows. 4. A method for producing a substrate for a microchip, which comprises forming a layer made of a silicon compound on the surface of a saturated cyclic polyolefin resin by using a dry coating method. 5. The method for manufacturing a microchip substrate according to claim 4, wherein the dry coating method is any one of a vacuum deposition method, a sputtering method and a CVD method.